Drug Testing and Analysis最新文献

Dried blood spots (DBS) have emerged as a promising complement, and in some settings, an alternative, to urine for anabolic androgenic steroid (AAS) testing, offering advantages such as minimal invasiveness, simplified storage, and transportation. This study evaluated two DBS collection devices—cellulose-based Capitainer-B50 and polymer-based Tasso-M20—and compared results with traditional urine analysis. Ten self-reported AAS users were recruited and provided matched urine and DBS samples. High agreement between the two DBS devices was observed, although Capitainer-B50 showed a slightly greater detection rate, likely due to a higher sample volume (50 μL vs. 17.5 μL) improved analyte recovery, and lower background noise. Notably, DBS enabled detection of testosterone use in all 10 participants, while urine testing missed two cases with naturally low urinary testosterone/epitestosterone (T/E) ratios (most likely UGT2B17 del/del carriers). Moreover, the differentiation between prescribed and illicit use of testosterone esters was also possible in DBS, but not in urine testing, while nandrolone detection in DBS was limited at low concentrations. The findings support DBS as a sensitive and practical tool for AAS detection and provide critical advantages in detecting doping with testosterone esters in individuals with prescribed testosterone therapy and in UGT2B17 deletion carriers.

RETRACTION: A. A. Ensafi and H. Karimi-Maleh, “ Voltammetric Determination of Isoproterenol Using Multiwall Carbon Nanotubes-Ionic Liquid Paste Electrode,” Drug Testing and Analysis 3, no. 5 (2011): 325–330, https://doi.org/10.1002/dta.232.

The above article, published online on 09 February 2011 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement of the journal Editor-in-Chief, Mario Thevis; and John Wiley & Sons Ltd. Following publication, concerns were raised by third parties regarding Figures 1A and 1B. The authors were unable to provide a satisfactory explanation and could not provide the original data. The retraction has been agreed because of concerns that the figures were duplicated and used to represent different materials. This affects the interpretation of the data and results presented.

The authors disagree with the decision to retract.

Tetrahydrocannabidiol (H4CBD) is an emerging semisynthetic cannabinoid, which has been known since 1940. Like hexahydrocannabinol (HHC), it is easily obtained by hydrogenation of available phytocannabinoids, in the case of H4CBD by hydrogenation of cannabidiol (CBD). H4CBD shows a weak affinity for the CB₁ receptor, but it is unclear if H4CBD shows psychoactive properties, as reports from users are divided. Only a few countries have placed H4CBD under their narcotic substance law, for example, France and Switzerland. The aim of this study was to identify human Phase I and II metabolites in urine as potential forensic targets. The H4CBD used for this study was bought from an online store and analyzed beforehand using GC–MS. The Phase I and II metabolites were identified using LC-HR-MS/MS and GC–MS after trimethylsilylation. The found H4CBD metabolites were carboxylated, hydroxylated, and bishydroxylated species and their glucuronides with hydroxylation and carboxylation positions on the alicyclic moiety and on the side chain. The tentatively identified metabolites were the carboxylic acids 5″-COOH-H4CBD and 7-COOH-H4CBD, the hydroxylated metabolites (1R,6R)-OH-H4CBD, (1R,6S)-OH-H4CBD, two epimers of 2″-OH-H4CBD, and both epimers of 7-OH-H4CBD. The identified bishydroxylated metabolites were side-chain hydroxylated derivatives of 7-OH-H4CBD. Various other hydroxylated metabolites were found, but their exact hydroxylation positions could not be determined. Some ESI+ spectra of the metabolites showed very unusual fragmentation patterns, like the loss of both oxygens from the resorcinol moiety with subsequent ring contraction and the appearance of radical cations for Phase II metabolites. These unusual patterns were noticed for H4CBD and its side-chain-altered metabolites.

Increasing oxygen transport through elevated hemoglobin concentration and red blood cell mass is a key objective of blood doping, commonly achieved via recombinant human erythropoietin (rHuEPO) administration or blood transfusions. While the Athlete Biological Passport (ABP) offers an effective indirect tool for detecting such manipulations, its sensitivity and specificity may be limited, particularly in cases involving microdoses or confounding physiological factors. To address these limitations, the identification of novel biomarkers that complement current ABP markers is essential.

This study presents a targeted metabolomics approach to discover candidate biomarkers of rHuEPO administration by analyzing polar metabolites in plasma and serum from two administration studies: one involving a single CERA injection, and the other using multiple doses of epoetin delta. Hydrophilic interaction chromatography hyphenated with tandem mass spectrometry enabled the selective and sensitive detection of a panel of polar endogenous metabolites.

Following data normalization and stringent quality control, generalized least squares models were applied to evidence temporal changes in metabolite signals. Among the most responsive and concordant markers across both studies were hypoxanthine and inosine, which showed significant and marked increases following rHuEPO administration. Notably, the relative increase of these metabolites coincided with the maximum in reticulocyte percentages, reflecting maximal erythropoietic activity. As intermediates in purine metabolism, their increases are likely tied to augmented purine turnover during red blood cell production. These findings suggest that hypoxanthine and inosine are promising candidate biomarkers to complement existing ABP parameters. However, further validation is required to confirm their reliability and applicability within the ABP framework.

Clenbuterol (Clb) is a β2-agonist drug included in the list of substances prohibited during and out of competition by the World Anti-Doping Agency (WADA-AMA). Several adverse analytical findings have been detected by accredited WADA laboratories, but athletes often claim that results are due to dietary contamination. In this contribution, bovine microsomal incubation and the excretion of bovine and human urinary metabolites of Clb were analyzed and compared using liquid chromatography electrospray Q-Exactive-Orbitrap mass spectrometry to determine differences in Clb metabolism. Urine samples were processed by solid-phase extraction prior to electrospray analysis in both the positive and negative ion modes. MS/MS experiments were obtained by parallel monitoring reaction (PRM) triggered by an inclusion ions list. The strategy for metabolite identification involved the search for typical biotransformation based on accurate mass shifts using diagnostic fragment ions from the parent drug. This approach successfully identified eight metabolites, including a novel N-methylated form of Clb, reported here for the first time. Additionally, four metabolites found exclusively in bovine urine offer significant potential for further research aimed at distinguishing unintentional doping.

Autologous blood transfusions (ABTs) are prohibited by the World Anti-Doping Agency (WADA), yet detecting autologous blood micro-transfusions (ABMTs) remains a challenge. Due to smaller transfused volumes, ABMTs cause attenuated biomarker changes, limiting detection sensitivity within the Athlete Biological Passport (ABP). This study assessed whether mRNA expression of 5-aminolevulinic acid synthase (ALAS2) and carbonic anhydrase 1 (CA1), measured from dried blood spots (DBS), could serve as sensitive biomarkers of ABMT. In a randomized, placebo-controlled design, 47 trained individuals (24 ♀; mean VO₂peak 56 ± 7 mL·min⁻¹·kg⁻¹) were allocated to an ABMT group (n = 23; ♀ = 12) or placebo group (n = 24; ♀ = 12). The ABMT group donated 450 mL of blood and received a 130 mL packed red blood cell reinfusion 4 weeks later. Blood sampling occurred regularly before and after both donation and reinfusion. ALAS2 and CA1 mRNA expression from DBS, and reticulocyte percentage (RET%) from venous blood, were analyzed. Following blood donation, ALAS2, CA1, and RET% increased by 270%, 200%, and 150%, respectively. However, no consistent group-level changes were observed after ABMT. Individualized analysis identified more outliers for ALAS2 than for CA1, and blinded interpretation of individual mRNA profiles achieved > 95% sensitivity and specificity for detecting ABMT. These findings suggest that ALAS2 mRNA expression, assessed via minimally invasive DBS sampling, is a promising biomarker for identifying ABMT. This approach may enhance current anti-doping strategies by improving sensitivity to small-volume autologous transfusions that evade detection through traditional ABP biomarkers.